Figure 8.

Effect of exogenous Runx1 expression on microglia activation. A, Western blot analysis of iNOS, amphiphysin II, and GAPDH expression in primary cultures of P3 mouse forebrain microglia, as indicated. Cells were not transduced (untran.) or transduced with adenovirus expressing GFP alone (GFP) or together with Runx1 (Runx1), followed by incubation in the absence or presence of LPS, as indicated. Cell lysates were collected and subjected to sequential Western blot analysis with anti-iNOS, anti-amphiphysin II (Amph-II), and anti-GAPDH antibodies. B, Double-label immunofluorescence analysis of GFP and iNOS expression in primary microglial cells that were either not transduced or transduced with adenovirus expressing GFP alone or together with Runx1, as indicated, followed by treatment with LPS. Where present, white arrows point to examples of cells expressing GFP but little or no iNOS, whereas yellow arrowheads point to examples of cells coexpressing GFP and iNOS. Notice how most cells transduced with GFP alone express iNOS, while most cells transduced with GFP + Runx1 express little or no iNOS. Scale bar, 50 μm.